Rab GTPases pp 195-205 | Cite as

Characterization of the Role Rab25 in Energy Metabolism and Cancer Using Extracellular Flux Analysis and Material Balance

  • Shreya Mitra
  • Jennifer Molina
  • Gordon B. Mills
  • Jennifer B. Dennison
Part of the Methods in Molecular Biology book series (MIMB, volume 1298)

Abstract

Rab25, by altering trafficking of critical cellular resources, influences cell metabolism and survival during stress conditions. Overall, perturbations in the vesicular trafficking machinery change cellular bioenergetics that can be directly measured in real time as Oxygen Consumption Rate, OCR (mitochondrial respiration) and Extracellular Acidification Rate, ECAR (glycolysis) by an extracellular flux analyzer (XF96, Seahorse Biosciences, MA). Additionally, overall turnover of glucose, lactate, as well as glutamine and glutamate can be measured biochemically using the YSI2900 Biochemistry Analyzer (YSI Incorporated, Life Sciences, OH). A combination of these two methods allows a precise and quantitative approach to interrogate the role of Rab25 as well as other Rab GTPases in central carbon energy metabolism.

Key words

Extracellular flux Glycolytic index Rab25 XF 96 extracellular flux analyzer YSI biochemistry analyzer Glucose Lactate Glutamine Breast cancer 

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Copyright information

© Springer Science+Business Media New York 2015

Authors and Affiliations

  • Shreya Mitra
    • 1
  • Jennifer Molina
    • 1
  • Gordon B. Mills
    • 1
  • Jennifer B. Dennison
    • 1
  1. 1.Department of Systems BiologyUT MD Anderson Cancer CenterHoustonUSA

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